2018-03-19T16:02:17Zhttp://journal.waocp.org/?_action=export&rf=summon&issue=69672018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Depression in Women with Breast Cancer: A Systematic Review of Cross-Sectional Studies in IranAzarJafariAmir HosseinGoudarzianMasoumehBagheri Nesami Objective: Breast cancer is one of the most common cancers in women and has more severe mental and emotional effects than other types. Depression as a mental disorder affects people’s mental well-being, physical symptoms, occupational performance, and finally quality of life. The aim of this study was to determine depression levels in Iranian women with breast cancer. Methods: A systematic review study was conducted in 2017. English and Persian databases (PubMed, SCOPUS, Web of Science, Google Scholar, SID, Magiran) were searched with key words such as Depression Or Depressive Disorders AND Women AND Breast Cancer OR Tumor OR Neoplasm OR Malignancy AND Iran. Inclusion criteria allowed for cross-sectional studies conducted in Iran (published in English or Persian language journals), studies that had key words in their keywords or their titles and standard instruments for measuring depression in patients. Of the 160 publications found, eight were selected after reviewing the title, abstract and full article. Results: Age of women with breast cancer in selected studies ranged from 43.8 (SD = 47.1) to 55.9 (SD = 14.6) years. Duration of cancer in most studies was about 1-2 years. In most studies, mild levels of depression for women with breast cancer were present. However, in one study it was stated that 69.4% of participants had serious levels of depression. Conclusions: There is increase in the risk of depression in women with breast cancer. Therefore, it seems necessary to plan preventive and therapeutic measures in order to improve the mental health and quality of life of the affected patients. depressionWomenBreast cancerMental HealthIran2018010117http://journal.waocp.org/article_55067_af8dbb928a6f409189d27ba567f852b8.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Attitude and Practice Regarding Breast Cancer Early Detection among Iranian Women: A Systematic ReviewMahinBadakhshAbbasBalouchiSafiyehTaheriSalehoddinBouyaSudabehAhmadidarrehsimaMohammadnaemAminifard Objectives: To determining attitudes and practice regarding breast cancer early detection techniques (breast self-examination (BSE), clinical breast examination (CBE) and mammography) among Iranian woman. Methods: International (PubMed, ISI, and Google Scholar) and national (SID and Magiran) databases were reviewed up to September 2017 to identify articles related to the attitudes and practices of Iranian women concerning breast cancer screening behavior with reference to BSE , CBE and mammography. The screening steps, analysis of quality of the studies and extraction of the papers were performed by two reviewers. Results: Of the 532 studies included initially, 21 performed on 10,521 people were considered eligible. Subjects with a positive attitude toward BSE in various studies were 13.5% to 94.0% with an average of 47.6%. Positive attitudes to CBE and mammography were found in 21.0% and 26.4%, respectively. Participant performance of BSE ranged from 2.6% to 84.7%, with an average of 21.9%. The respective figures for CBE and mammography were 15.8% and 16.7%. Conclusion: Considering the poor performance and low rates for positive attitudes, it is suggested that educational programs should be conducted across the country. Breast Self-ExaminationEarly Detection of CancerAttitudeIranSystematic reviews20180101916http://journal.waocp.org/article_55097_2a884a7e06bf5e5a0b83839c19010ad6.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191More Reasonable Animal Model for Study the Effect of Pneumoperitoneum on Abdominal Tumor CellsLvQijunDuJiangWangChongshu Background: Many animal experimental studies showed that abdominal tumor cells will be widely spread during laparoscopic treatment and grow into metastases. These results are different from clinical observations. There is a hypothesis that too much tumor cells was injected in the animals lead to the results of theses bias. We aim to learn the difference of abdominal cavity volume between human body and the nude mice and to determine reasonable amount of tumor cells in the animal experiments. Methods: The insufflated CO2 volume which represents the capacity of the abdominal cavity was recorded during laparoscopic process in 212 patients and 20 nude mice respectively, the relative volume of nude mice and human body was calculated.Based on data from the literature and this study , the amount of tumor cells in the animal experiments was determined.According to these data, we set up a new animal model and a traditional one respectively,and compared the rate of successful modeling and tumor formation between two animal models. Results: The intraperitoneal volumes of humans and nude mice were 3.01±0.36 L and 0.011±0.001 L respectively.The number of tumor cells that be uesd in animal should be approximately 0.26×105 in terms of known data in human beings.Compared with the traditional animal model which formed a large number of intraperitoneal tumor metastasis, the new animal model was shows more moderately,and the rate of successful modeling was similar. Conclusion:In animal experiments, to simulate the clinical situation, about 0.26×105 tumor cells should be inject in peritoneal cavity of the nude mice. human bodypneumoperitoneumtumor cellsanimal models201801011720http://journal.waocp.org/article_54214_336538eea9da2b8f14e9c97b71b14857.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191The Economic Burden of Metastatic Castration Resistant Prostate Cancer and Skeletal Related Events in Japanese University HospitalsTakefumiSatohDianneLedesmaNariakiYoshiharaObjective: Although androgen deprivation therapy (ADT) has improved the survival and quality of life of patientswith prostate cancer, resistance to treatment inevitably results in transition to a castration resistant state (CRPC) and, inadvanced cases, bone metastasis, leading to skeletal related events (SRE). In order to understand the current burden onpatients in Japan, there is a need to estimate the healthcare costs of CRPC treatment in current clinical practice. Methods:This retrospective observational cohort study utilized claims data from 13 national university hospitals through thePlatform for Clinical Information Statistical Analysis database. Extracted data included the use of diagnostic tests, thefrequency and cost of hospitalizations and outpatient visits, and medication costs, using values from the Healthcare FeeSystem and the National Health Insurance Drug Price List relative to each observed year. Results: Data were collectedfrom 4001 patients with CRPC, 97% of whom had undergone ADT. Between 2005 and 2016, the mean annualizeddirect medical cost per patient was ¥739,147 (US$7060), of which 91% was related to medication, 4.8% to laboratoryand imaging, 4.1% to radiotherapy, and 0.1% to surgery. A total of 771 (19%) of the 4001 CRPC patients experiencedan SRE. Resource utilization was significantly higher (pMetastatic castration resistant prostate cancerskeletal related eventsburdenJapan201801012126http://journal.waocp.org/article_57911_4067ba3cdffbf15b7a8c2447dc331ed3.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Prognostic Value of IL-10 and Its Relationship with Disease Stage in Iranian Patients with Multiple MyelomaRaminShekarrizGhasemJanbabaiSaeedAbedian Kenari Background: Several studies have demonstrated roles of interleukins in the pathogenesis of multiple myeloma (MM). Objective: Here we considered correlations among serum levels of IL-10, stage of disease and clinical laboratory disease markers in Iranian MM patients to investigate whether the interleukin might have prognostic significance. Materials and Methods: In this cross-sectional study, a total of 60 subjects (40 patients and 20 controls) were recruited. After preliminary laboratory tests, disease stage was evaluated and serum levels of IL-10 were measured using an enzyme-linked immunosorbent assay (ELISA). Results: The mean concentration of serum IL-10 in patients (2.39±0.82 ng/ ml) was significantly higher (pmultiple myelomaIL-10disease stagesplasma cells201801012732http://journal.waocp.org/article_54210_edf7c8c3825ee29463d459051b37dc00.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Serum MicroRNA-21 Negatively Relates to Expression of Programmed Cell Death-4 in Patients with Epithelial Ovarian CancerEnas HMahmoudAmalFawzyReham AA Elshimy Background: Ovarian cancer is the third most common cancer of the female genital tract and the leading cause of cancer death associated with gynecologic tumors. MicroRNAs regulate at least 60% of human genes, including tumor suppressor genes and oncogenes and, thereby, can affect cancer risk. Aim of the work: We aimed to assess any diagnostic role for serum miR-21 as a biomarker in human ovarian cancer and to study relations with programmed cell death-4 (PDCD4), one of its target proteins, hoping to help explain heterogeneity of this cancer type and facilitate stratification of regimens for therapy. Subjects and Methods: A total of 60 newly diagnosed ovarian cancer cases and 30 apparently healthy females were recruited. Serum microRNA-21 levels were measured by TaqMan- Real time PCR assay and PDCD4 by ELISA. Results: Significant over-expression of serum miR-21 and lower serum PDCD4 levels were observed in ovarian cancer patients as compared to the control group. A statistically significant inverse correlation was also evident between miR-21 and PDCD4. However, no significant links were noted observed between miR-21 and tumor grade, stage or histopathological type. Conclusion: The present work showed significantly up-regulation of serum miR21 in the recruited group of patients and a significant inverse relation association between miR-21and PDCD4. These findings suggest that miR-21 may be used as a diagnostic biomarker for human ovarian cancer. Key word: ovarian cancermiR-21PDCD4201801013338http://journal.waocp.org/article_55092_311c977aee53a45b69e7b98170d8625d.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Long-term Results of Adjuvant Imatinib Treatment for Localized Gastrointestinal Stromal Tumors after SurgeryMozaffarAznabSayed MojtabaAkhmadi Objective: Despite the development of two significant classifications for recurrence risk evaluation among patients engaged with gastrointestinal stromal tumor and corresponding treatment criteria, recurrence happens in a number of the patients who were once classified as ineligible for treatment and hence removed from treatment program. As such, the aim of the present study is to increase the number of patients recognized as eligible for treatment, so as to further reduce recurrence rate of this disease. Materials and Methods: A total of 26 patients from Ilam, Kermanshah, Lorestan, Kurdistan, and some parts of Hamedan, entered this study from 2006 until 2016. The western provinces included have similar socioeconomical conditions. Inclusion criteria were operable tumors confirmed radiologically with a gross size larger than 3 centimeters regardless of the mitosis rate in microscopic power fields, tumor location, or presence of peritoneal involvement during the surgery. Imatinib capsules were administered daily at 400 mg for 3 years. The patients were followed up every 3 months by radiology, ultrasonography, biochemical assessment, and clinical examination. Results and Conclusions: The overall survival after 10-years follow up was 100%, while 5-year survival without relapse was 95%. Mean overall survival was 106 months, and only one patient who had limited peritoneal involvement experienced relapse and he is still alive after 2 years. The drug was well tolerated and no significant side effects were observed. GISTImatinibTarget Therapy2018010139432018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Fas-Antisense Long Noncoding RNA and Acute Myeloid Leukemia: Is There any Relation?ArezouSayadAbbasHajifathaliAmir AliHamidiehFarbodEsfandiMohammadTaheri In recent years, lncRNAs have been considered as potential predictive biomarkers for prognosis of different human cancers. One example is the FAS antisense RNA 1 (FAS-AS1) located in the 10q23.31 region which is transcribed from the opposite strand of the FAS gene. FAS has an important role in regulation of apoptotic pathways and there is an inverse correlation between FAS-AS1 expression level and production of the soluble form of Fas, so that it might have potential as a therapeutic target to improve chemotherapy effectiveness. In the present study we therefore evaluated FAS-AS1 expression in blood samples of de novo AML patients and healthy controls using real-time quantitative reverse transcription-PCR (qRT-PCR). Our results indicated that the expression level of FAS-AS1 lncRNA demonstrated no significant difference between AML patients and healthy individuals. We conclude from the obtained data that FAS-AS1 is not an informative and reliable biomarker for AML diagnosis, although our results need to be confirmed in further studies. Acute myeloid leukemialong non-coding RNAFAS-AS1201801014548http://journal.waocp.org/article_55068_af20acbcae903bd2336fdf14c80eb8c8.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Curcumin Analog Pentagamavunon-1 (PGV-1) Sensitizes Widr Cells to 5-Fluorouracil through Inhibition of NF-κB ActivationEdyMeiyantoEndah PujiSeptisetyaniYonika ArumLarasatiMasashiKawaichi Cell cycle regulation and the NF-κB pathway in cancer cells are important in mediating resistance to 5-Fluorouracil (5-FU). Pentagamavunon-1 (PGV-1), a curcumin analog, is known to exhibit stronger growth inhibitory effects than curcumin itself in several cancer cells. In this study, we evaluated the potency of PGV-1 in combination with 5-FU in WiDr colon cancer cells. In MTT assays, PGV-1 did not only exhibit stronger growth inhibitory effects than both 5-FU and curcumin, but also enhanced the cytotoxicity of 5-FU. Flow cytometry demonstrated that single treatments with PGV-1 and 5-FU resulted in different effects on cell cycle profiles. PGV-1 induced G2/M arrest while 5-FU caused S-phase arrest at low concentration (1 μM) and G1-phase arrest at high concentration (100 μM). Interestingly, the combination of 5-FU and PGV-1 enhanced cell accumulation in S-phase. Although a single treatment with either 5-FU or PGV-1 increased cyclin D1 at the protein level, the combination treatment resulted in significant suppression. In addition, PGV-1 inhibited activation of NF-κB and suppressed the expression of cyclooxygenase-2, an NF-κB downstream protein. In conclusion, PGV-1 increased the cytotoxic effect of 5-FU on WiDr cells through inhibition of NF-κB activation. Cell cycle5-fluorouracilNF-BPGV-1WiDr cells201801014956http://journal.waocp.org/article_55070_eef5d96d292756f625562a06a6b39680.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191The Effect of Mammary Extracellular Matrix in Controlling Oral and Mammary Cancer CellsSivapriyaPavuluriJulie ASharpChristopheLefevreKevin RNicholas Extracellular matrix (ECM) plays an important role in the normal physiology of tissues and progression to disease. Earlier studies and our external microarray data analysis indicated that mammary matrix from involuting tissue showed upregulation of genes involved in ECM remodeling. The present study examines the fate of mammary and oral cancer cells grown in the ECM from lactating mammary gland. Our findings show that non-tumorigenic cells, MCF10A and DOK cells did not proliferate but the tumorigenic and metastatic cells, SCC25 and MDA-MB-231, underwent apoptosis when grown on mammary ECM isolated from lactating mice. In addition, the cytokinesis marker, CEP55, was repressed in the oral and breast cancer cells. In contrast, these cells proliferated normally on mammary ECM isolated from mice undergoing involution. External microarray data analysis of mammary tissue further revealed over expression (~16 fold) of QSOX1 gene, which promotes cellular quiescence, in lactating mammary gland. A recent study has indicated that QSOX1 overexpression in breast cancer cells led to reduced proliferation and tumorigenic properties. This extracellular protein in mammary ECM may be responsible for reduced cellular proliferation. The present study has shown that ECM from lactating mammary gland can regulate signals to oral and breast cancer cells to halt cell division. This preliminary observation provided insights into the potential role of ECM factors present in lactating mammary gland as therapeutic targets to control cancer cell division. This preliminary study is an attempt to understand not only the requirement of ECM remodeling factors essential for the growth and survival of cancer cells but also the factors present in the lactation matrix that simultaneously halts cell division and selectively inhibits the growth of cancer cells. CancerExtracellularmatrixLactationInvolution201801015763http://journal.waocp.org/article_55060_5b482a11eb31663e71c4825148c96e2c.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Health-Related Quality of Life before and after Surgical Resection of Hepatocellular Carcinoma: A Prospective StudyChong-ChiChiuKing-TehLeeJhi-JoungWangDing-PingSunHao-HsienLeeHon-YiShi Background and Objectives: This large-scale prospective cohort study of a Taiwan population applied generalized estimating equations (GEEs) to evaluate changing trends in health-related quality of life (HRQoL) and to compare predictors of HRQoL before and after surgical resection of hepatocellular carcinoma (HCC) performed during 2011-2014. Materials and Methods: The Short Form-36 Health Survey (SF-36) and Functional Assessment of Cancer Therapy-Hepatobiliary were used in a preoperative assessment and in 3- and 6-month postoperative assessments of 332 HCC patients. Results: The HRQoL was signficantly (pHepatocellular carcinomahealth-related quality of lifeSF-36FACT-Hep201801016572http://journal.waocp.org/article_55119_bd0de6ea5294e03869fe7a47001fc6b7.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Pattern of Failure with Locally Advanced Cervical Cancer– A Retrospective Audit and Analysis of Contributory FactorsAnisBandyopadhyayUpasanaMukherjeeSandipGhoshSauravGhoshShyamal KumarSarkar Background: The majority of the global burden of cervical cancer is affecting developing countries. Despite improvement in treatment of patients presenting at a locally advanced stage, approximately 50% experience recurrence within the 1st two years. This study was conducted to analyse contributory factors for recurrence within 24 months. Methods: The present retrospective study was undertaken to analyse factors affecting recurrence, type of failure and the follow up pattern of patients who completed treatment with a minimum follow-up period of 6 months during the study period of 5 years. Results: Out of 323 patients included in the study, 112 (34.7%) presented with recurrence within the follow-up period. The stage and histology had a significant impact on disease free survival (DFS). Of those who were followed-up regularly, recurrence was observed in 28.7% with a DFS of 81.3 months, in contrast to the 48. 5% patients with a DFS of 45.0 months for whom follow-up was irregular. The failure pattern was mostly in the form of nodal recurrence (61%). On univariate analysis, treatment time, EBRT and ICBT gap and mean EQD2 point A were found to associated with a better outcome in terms of 2yr DFS. On Cox regression analysis, stage, histology, treatment gap (HR-0.48) and follow up pattern (HR-0.24) retained their effects on survival. Point A dose was higher in patients without recurrence (P value 0.000) unlike other assymmetric parameters. Conclusion: Apart from point A cumulative dose (mean EQD2), stage, histology and treatment gap were the factors that affected early local failure. An interesting result was that follow-up pattern had a significant impact on DFS period. : Retrospective auditlocally advanced cervical cancerearly local recurrence201801017379http://journal.waocp.org/article_53943_fb317c1b56cc4982e3fae7ebfd20d01c.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Differential Response of B Cells to an Immunogen, a Mitogen and a Chemical Carcinogen in a Mouse Model SystemNimishaSaxenaAmar PreetKaurNimai ChandChandra Background: B cells are specific antibody generating cells which respond to foreign intruders in the circulation. The purpose of this study was to compare the relative immunogenic potentials of three well established agent types viz. an immunogen, a mitogen and a carcinogen, by following B cell responses to their presence in a mouse model system. Methods: Mice were treated with tetanus toxoid (immunogen), poke weed mitogen (typical mitogen), and benzo-α- pyrene (carcinogen) and generated B cell populations were determined in isolated splenic lymphocytes (splenocytes) by flow cytometry using specific anti-B cell marker antibodies. Flow cytometric estimation of LDL receptor (LDLR) expression, along with associated B cell markers, was also conducted. Kit based estimation of serum IgG, western blotting for LDLR estimation on total splenocytes and spectrometry for cholesterol and serum protein estimation were further undertaken. Student’s T-tests and one way ANOVA followed by the Bonferroni method were employed for statistical analysis. Results: The mitogen was found to better stimulate B cell marker expression than the immunogen, although the latter was more effective at inducing antibody production. The chemical carcinogen benzo-α-pyrene at low concentration acted potentially like a mitogen but almost zero immunity was apparent at a carcinogenic dose, with a low profile for LDLR expression and intracellular cholesterol. Conclusion: The findings in our study demonstrate an impact of concentration of BaP on generation of humoral immunity. Probably by immunosuppression through restriction of B-cell populations and associated antibodies, benzo-α-pyrene may exerts carcinogenicity. The level of cholesterol was found to be a pivotal target. Key words: B cellsIgGLdl receptorCholesterolBenzo-alpha-pyrene201801018190http://journal.waocp.org/article_54206_f83de5c25149f1938187642ed98fe432.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Association of ARID5B Genetic Variants with Risk of Childhood B Cell Precursor Acute Lymphoblastic Leukaemia in LatviaMadaraKreileDmitrijsRotsAgneseZarinaLindaRautiainenZelmaVisnevska-PrecinieceZhannaKovalovaLindaGailite Background: Acute lymphoblastic leukaemia (ALL) is the most common malignancy in childhood. Despite numerous investigations very little is still known about its aetiology. However, in one genome wide association study conducted to identify the possible genetic risk factors, two allelic variations rs10821936 and rs10994982 in the 3rd intron of the ARID5B gene were identified as possible ALL risk alleles. Association between ARID5B gene variants and ALL risk was also been confirmed for different ethnic groups. Materials and Methods: Eight genetic variants in the gene ARID5B were genotyped - rs10994982, rs7908445, rs7923074, rs10821936, rs10821937, rs7896246, rs10821938 and rs7089424 in 77 ALL patients in remission and in 122 age and gender matched controls; parental samples were also genotyped in 50 cases. Results: Six out of the eight (rs7908445, rs7923074, rs10821936, rs10821937, rs7896246 and rs7089424) analysed allelic variations were identified in the case-control analysis as statistically significant risk alleles for ALL development. In the family study and using hybrid analysis, all allelic variations were significantly associated with ALL. During the study, risk haplotype was identified rs10994982/rs7908445/rs7923074/ rs10821936/ rs10821937/rs7896246/rs10821938/rs7089424 – ATACCAAG – with a frequency in cases of 0.17 and in the control group at 0.29 (chi square = 6.69, p value = 0.009). In the family association study the same haplotype showed statistical significance (chi squared = 10.3, p value = 0.001). Conclusions: Results of the study replicate and extend previously published findings for ARID5B localized allelic variants, but do not explain the mechanism of action related to the pathogenesis of ALL. Childhood ALLARID5BSusceptibilityLatviagenetic variants201801019195http://journal.waocp.org/article_54212_8319cd7ebc44bd4a0ad9c0bd19e65381.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Anti-ROR1 scFv-EndoG as a Novel Anti-Cancer Therapeutic DrugPeymanBemaniMozafarMohammadiAliHakakian Aim: Immunotoxins are proteins that consist of an antibody fragment linked to a toxin, used as agents for targeted therapy of cancers. Although the most potent immunotoxins are made from bacterial and plant toxins, obstacles which contribute to poor responses are immunogenicity in patients and rapid development of neutralizing antibodies. In the present study we proposed a new therapeutic immunotoxin for targeted cancer therapy of ROR1 expressing cancers: an anti ROR1 single chain fragment variable antibody (scFv)-endonuclease G (anti ROR1 scFv-EndoG). Methods: The three-dimensional structure of anti ROR1 scFv-EndoG protein was modeled and structure validation tools were employed to confirm the accuracy and reliability of the developed model. In addition, stability and integrity of the model were assessed by molecular dynamic (MD) simulation. Results: All results suggested the protein model to be acceptable and of good quality. Conclusions: Anti-ROR1 scFv-EndoG would be expected to bind to the ROR1 extracellular domain by its scFv portion and selectively deliver non-immunogenic human endonuclease G enzyme as an end-stage apoptosis molecule into ROR1-expressing cancer cells and lead rapidly to apoptosis. We believe that anti ROR1 and other anti-tumor antigen scFv-EndoG forms may be helpful for cancer therapy. Cancer therapyimmunoconjugatescFvEndoGapoptosis2018010197102http://journal.waocp.org/article_54001_de992e4e0d7446502d6cce20f6df3c78.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Mitochondrial Effects of Teucrium Polium and Prosopis Farcta Extracts in Colorectal Cancer CellsForouzanKhodaeiKiyanooshAhmadiHamzeKiyaniMahmoudHashemitabarMohsenRezaei Background: Teucrium Polium and Prosopis Farcta have been traditionally employed in cancer treatment. In this study we evaluated the effects of methanolic extracts of these two plants in HT-29 cells. Methods: IC50s of extracts were obtained via MTT assay and the levels of ROS production, cell death, collapse of mitochondrial membrane potential and Sirt3 enzyme activity were determined. Results: After 48 hours exposure, IC50s for Teucrium and Prosopis extracts were 3 and 2μg/ml, respectively. Extracts induced higher ROS production after 6 hours than after 12 hours. Mitochondrial membrane potential collapse and cell death rate were also increased; Teucrium caused greater cell death than Prosopis. Extracts from both plants increased Sirt3 activity in its normal form, but only Teucrium extract caused a significant increase in activity of Sirt3 enzyme isolated from cancer cells. Conclusion: Teucrium and Prosopis extracts exert anticancer activity via mitochondrial alterations, as exemplified by increased ROS levels, Sirt3 activity and cell death in HT-29 colorectal cancer cells. Teucrium PoliumProsopis FarctaSirt3ROSapoptosis20180101103109http://journal.waocp.org/article_54325_528d05dfcaadf04cb024e4652c173109.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Epidermal Growth Factor Receptor Mutations in Lung Adenocarcinomas: A Single Center Study from IranAliBasiFloraKhalediMohammad HadiKarbalaie NiyaHamidRezvaniNasserRakhshani Introduction: Lung cancer is the fifth leading tumor in Iran, and while its incidence remains relatively low, it has been increasing steadily. Targeted therapies have brought new hope to patients with non small cell lung cancer (NSCLC). The epidermal growth factor receptor (EGFR) gene is the prototype member of the type I receptor tyrosine kinase (TK) family and plays a pivotal role in cell proliferation and differentiation. Studies from Asian countries have revealed a higher frequency of EGFR mutations than in the West. The aim of this study was to measure the frequency and type of EGFR mutations in a group of Iranian patients with lung adenocarcinomas. Methods: Formalin fixed paraffin embedded (FFPE) lung adenocarcinoma tissues from 103 Iranian patients were sequentially tested for EGFR mutations by the polymerase chain reaction (PCR) followed by direct nucleotide sequencing of exons 18, 19, 20, and 21. Patient’s demographics and other clinical details were obtained from the medical records of hospitals affiliated to Iran University of Medical Sciences, Tehran, Iran. Statistical analyses were performed with SPSS v.20. Results: EGFR mutations were detected in 25/103 (24.3%) patients. The most frequent was an exon 21 point mutation (L858R) (15 patients; 60%), followed by one in exon 19 (10 patients; 40%). The frequency of EGFR mutations in never-smoker patients was significantly higher than in smokers (68% versus 32%; p < 0. 01). Conclusion: EGFR mutation frequency is higher than in the West but lower than in East Asian and almost equal to reported rates for Indian and North African populations. Smoking is negatively associated with EGFR mutations in Iranian lung adenocarcinomas. non-small cell lung cancerepidermal growth factor receptormutationreceptor tyrosine kinase20180101111114http://journal.waocp.org/article_55095_30a01f2133ee33c6dec06e6faff6e7fb.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Compliance with National Guidelines on the Treatment of Stage II–IVB Nasopharyngeal Carcinoma in a Regional Cancer Center of Southern ChinaJia-xiangYeXiaLiangJianWeiJingZhouYuLiaoYu-LeiLuXia-QuanTangAn-YuWangYongTang Objective: It is unknown whether the treatment provided to patients with stage II-IVB NPC in southern China adheres to the 2015 NCCN guidelines. Consequently, a retrospective analysis was conducted, in order to evaluate the compliance with NCCN guidelines and identify the areas for improvement. Methods: The present study was a retrospective study that included patients with stage II-IVB NPC in southern China during the period 2013 and 2014. The treatment regimens were compared with the 2015 NCCN guidelines in order to identify potential noncompliance regarding the treatment for stage II–IVB NPC. The statistical analyses included descriptive statistics, univariate and/or multivariate analysis using SPSS version 16.0.0. Results: A total of 215 patients, including 166 men (77.21%) and 49 women (22.79%), were involved in the analysis. Although the overall rate of noncompliance with the NCCN recommendations was 23.26%, the noncompliance rate of concurrent chemoradiation (CCRT), induction of chemotherapy (IC) followed by CCRT and CCRT followed by adjuvant chemotherapy (AC) was 7.02%, 39.76% and 50.00%, respectively. Univariate analysis indicated that NCCN noncompliance regarding the treatment for stage II-IVB NPC did not exhibit a significant correlation with the parameters age, gender, insurance status, education profile, first clinic department, careers, comorbidities and overall clinical stage, but it indicated a significant association with the therapeutic schedule (Pnasopharyngeal neoplasmscomplianceguideline20180101115120http://journal.waocp.org/article_55063_491b5cd4d2ff6d111090f3a1efeab907.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Beliefs and Behavior of Saudi Women in the University of Tabuk Toward Breast Self Examination PracticeAnalitaGonzalesMohammadAlzaatrehMohammadMariAbdulmoneamA SalehAladeenAlloubani Background: Breast cancer is one of the most frequent types of malignancy worldwide, Breast Self Exam (BSE) is considered as a simple method to screen and detect breast cancer, then early beginning of treatment and enhancing survival rates. Aim: To Identify the health beliefs about breast Self-Examination and its relationships with the frequency of BSE among the women in the University of Tabuk at Saudi Arabia. Methods: Descriptive, cross-sectional correlational design was used; Champion Health Beliefs Model (CHBM) was utilized to assess health beliefs among 400 women who answered a self-administered questionnaire. Results: Among the 400 respondents, almost all the sample (382,95.5%) heard about BSE. However, only (7.8%) practiced BSE regularly each month in the past year, and (9%) is intended to perform BSE monthly in future. There was a positive relationship between performing BSE last year and the beliefs of susceptibility and confidence. While, intention to perform BSE in the future was significantly correlated to seriousness and confidence. Perceived barriers were negatively related to BSE performance in last 12 months and in future. Conclusion: Health beliefs affect the behavior of women in practicing BSE. Confidence was related Breast self-exam (BSE)Breast cancerHealth beliefs model (HBM)Saudi Arabia20180101121126http://journal.waocp.org/article_54215_55ce389f00d5cece1d4529a10c7adf56.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Hemostatic Abnormalities in Multiple Myeloma PatientsAartiGogiaMeeraSikkaSatenderSharmaUshaRusia Background: Multiple myeloma (MM) is a neoplastic plasma cell disorder characterized by clonal proliferation of plasma cells in the bone marrow. Diverse hemostatic abnormalities have been reported in patients with myeloma which predispose to bleeding and also thrombosis. Methods: Complete blood count, biochemical parameters and parameters of hemostasis i.e. platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), factor VIII assay results, plasma fibrinogen, D-dimer and lupus anticoagulant, were assessed in 29 MM patients and 30 age matched controls. Results: The most frequent abnormal screening parameter was APTT. Of the six indicative of a bleeding tendency i.e. thrombocytopenia, prolonged PT, APTT, TT, reduced plasma fibrinogen and factor VIII, at least one was abnormal in 8 (27.6%) patients. Of the four prothrombotic markers, lupus anticoagulant, D-dimer, elevated factor VIII and plasma fibrinogen, one or more marker was present in 24 (82.7%). D-dimer was the most common prothrombotic marker, being elevated in 22 (75.9%) patients. One or more laboratory parameter of hemostasis was abnormal in all 29 (100%) patients. Though thrombotic complications are reported to be less frequent as compared to hemorrhagic manifestations, one or more marker of thrombosis was present in 24 (82.7%) patients. Conclusion: This study provided laboratory evidence of hemostatic dysfunction which may be associated with thrombotic or bleeding complications at diagnosis in all MM patients. Hence, screening for these abnormalities at the time of diagnosis should help improved prognosis in such cases. multiple myelomaHemostatic abnormalitiesThrombosisBleeding20180101127130http://journal.waocp.org/article_54081_67f680ac6d873d8ac4c4675f39ae5d8b.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Eupatorin and Salvigenin Potentiate Doxorubicin-Induced Apoptosis and Cell Cycle Arrest in HT-29 and SW948 Human Colon Cancer CellsNazaninNamazi SarvestaniHouriSepehriLadanDelphiMahdiMoridi Farimani Background: Cancer persists as one of the world’s most pressing maladies. Notable points about chemotherapy are drug side effects which are almost universally encountered. Emerging knowledge focusing on mechanisms of toxicity due to chemotherapy has led to characterization of novel methods, including the exploitation of natural compounds, in combination therapies. Flavonoids are natural polyphenolic compounds that play protective roles against tumor cell development. The focus of this study was apoptotic effects of two flavonoids, eupatorin and salvigenin, in combination with doxorubicin on a cellular model of colon cancer. Method: Upon establishing a non-effective dose of doxorubicin, and effective doses of eupatorin (100μM) and salvigenin (150μM) via MTT, morphological features of apoptosis were distinguished using DAPI staining and cell cycle blockage in the sub-G1 phase. Apoptosis was determined by annexin/ PI and western blotting. ROS levels and MMP were measured to show any role of mitochondria in apoptosis. Results: Co-administration of flavonoids with doxorubicin induced apoptosis via the mitochondrial pathway as mitochondrial membrane potential and ROS production were changed. Annexin/PI analysis demonstrated that apoptosis frequency was increased with the combination treatments in colon cancer cells. Finally, the combination of these flavonoids with doxorubicin increased the Bax/Bcl-2 ratio, caspase-3 expression and PARP cleavage. Conclusion: Combination of flavonoids with doxorubicin induces apoptosis and enhances effect on cancer cells which might allow amelioration of side effects by dose lowering. doxorubicinEupatorinHT-29SalvigeninSW94820180101131139http://journal.waocp.org/article_55093_65e835eb05feb902ee084ba42b8d8ae1.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Significance of DNA Replication Licensing Proteins (MCM2, MCM5 and CDC6), p16 and p63 as Markers of Premalignant Lesions of the Uterine Cervix: Its Usefulness to Predict Malignant PotentialVNSarithaVSVeenaKMJagathnath KrishnanTharaSomanathanKSujathan Cervical cancer continues to be a leading cancer among women in many parts of the world. Nation-wide screening with the Pap smear has not been implemented in India due to the lack of adequately trained cytologists. Identification of biomarkers to predict malignant potential of the identified low risk lesions is essential to avoid excessive retesting and follow up. The current study analyzed the expression patterns of DNA replication licensing proteins, proliferation inhibitor protein p16INK4A and tumor suppresser protein p63 in cervical tissues and smears to assess the ability of these proteins to predict progression. Methods: Cervical smears and corresponding tissues were immunostained using mouse monoclonal antibodies against MCM2, MCM5, CDC6, p16 and p63. Smears were treated with a non-ionic surfactant sodium deoxycholate prior to immuno-cytochemistry. The standard ABC method of immunohistochemistry was performed using DAB as the chromogen. The immunostained samples were scored on a 0-3+ scale and staining patterns of smears were compared with those of tissue sections. Sensitivity and specificity for each of these markers were calculated taking histopathology as the gold standard. Result: All the markers were positive in malignant and dysplastic cells. MCM protein expression was found to be up-regulated in LSIL, HSIL and in malignancies to a greater extent than p16 as well as p63. CDC6 protein was preferentially expressed in high grade lesions and in invasive squamous cell carcinomas. A progressive increase in the expression of DNA replication licensing proteins in accordance with the grades of cervical intraepithelial lesion suggests these markers as significant to predict malignant potential of low grade lesions in cervical smears. Conclusion: MCMs and CDC6 can be applied as biomarkers to predict malignant potential of low grade lesions identified in screening programmes and retesting Cervical CancerHPVbiomarkerscervical intraepithelial neoplasia20180101141148http://journal.waocp.org/article_55079_ade673a9e2da0f9da6d2e6914067ff6f.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Thai Water Lily Extract Induces B16 Melanoma Cell Apoptosis and Inhibits Cellular Invasion Through the Role of Cellular OxidantsParichayaAimvijarnSarawootPalipochSeijiOkadaPrasitSuwannalert Melanoma is a cancer that is associated with a high capacity of invasion. Oxidative stress is recognized as cancer growth and progression. The phytochemical pigments of natural products show either anti-oxidant or pro-oxidant activity from the redox system. In addition, the phytophenolics also prevent cancer cell proliferation and progression. Objective: This study aims to investigate the effects of Thai water lily on cell apoptosis and cellular invasion through the role of cellular oxidants in B16 melanoma cells. Methods: The cytotoxicity and cell apoptosis of Thai water lily extract treating B16 cells were performed by using the MTT and Annexin V/PI-flow cytometry methods, respectively. In addition, cellular oxidants and cancer cell invasion were also obtained by using DCFH-DA and Boyden chamber assays, respectively. Results: Thai water lily, Nymphaea stellate extract was shown to be markedly toxic to B16 melanoma cells with IC50 = 814 μg/ml. The extract at 800 and 1,000 μg/ml demonstrated pro-oxidant activity relating to the cell apoptosis. The low concentrations of the extract at 200 and 400 μg/ml showed the anti-oxidant function associated with the inhibitory effect of melanoma cell invasion. Conclusion: Thai water lily extract may play an important role in bioactive work as a chemo preventive agent on the modulation of cellular oxidative stress-induced apoptosis and suppressed cancer cell invasion. Thai water lilyMelanoma cellsapoptosisInvasionoxidative stress20180101149153http://journal.waocp.org/article_53860_2fa07b435999cca6e0607178b20636e3.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Knowledge and Practices Related to Screening for Breast Cancer among Women in Delhi, IndiaNehaDahiyaSauravBasuMegha ChandraSinghSuneelaGargRajeshKumarCharuKohli Background: Breast cancer is a major public health problem globally. The ongoing epidemiological, socio-cultural and demographic transition by accentuating the associated risk factors has disproportionately increased the incidence of breast cancer cases and resulting mortality in developing countries like India. Early diagnosis with rapid initiation of treatment reduces breast cancer mortality. Therefore awareness of breast cancer risk and a willingness to undergo screening are essential. The objective of the present study was to assess the knowledge and practices relating to screening for breast cancer among women in Delhi. Methods: Data were obtained from 222 adult women using a pretested self-administered questionnaire. Results: Rates for knowledge of known risk factors of breast cancer were: family history of breast cancer, 59.5%; smoking, 57.7%; old age, 56.3%; lack of physical exercise, 51.9%; lack of breastfeeding, 48.2%; late menopause, 37.4%; and early menarche, 34.7%. Women who were aged < 30 and those who were unmarried registered significantly higher knowledge scores (p ≤ 0.01). Breast self-examination (BSE) was regularly practiced at-least once a month by 41.4% of the participants. Some 48% knew mammography has a role in the early detection of breast cancer. Since almost three-fourths of the participants believed BSE could help in early diagnosis of breast cancer, which is not supported by evidence, future studies should explore the consequences of promoting BSE at the potential expense of screening mammography. Conclusion: Our findings highlight the need for awareness generation among adult women regarding risk factors and methods for early detection of breast cancer. Breast cancerknowledgeRisk factors20180101155159http://journal.waocp.org/article_54327_462e9607769bbbdae0d5be50af6b9ccb.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Molecular Subtypes of Indonesian Breast Carcinomas - Lack of Association with Patient Age and Tumor SizeYeniRahmawatiYunitaSetyawatiIrianiwatiWidodoAhmadGhozaliDewajaniPurnomosari Objective: Breast carcinoma (BC) is a heterogeneous disease that exhibits variation in biological behaviour, prognosis and response to therapy. Molecular classification is generally into Luminal A, Luminal B, HER2+ and triple negative/basal-like, depending on receptor characteristics. Clinical factors that determined the BC prognosis are age and tumor size. Since information on molecular subtypes of Indonesian BCs is limited, the present study was conducted, with attention to subtypes in relation to age and tumor size. Methods: A retrospective cross-sectional study of 247 paraffin-embedded samples of invasive BC from Dr. Sardjito General Hospital Yogyakarta in the year 2012- 2015 was performed. Immunohistochemical staining using anti- ER, PR, HER2, Ki-67 and CK 5/6 antibodies was applied to classify molecular subtypes. Associations with age and tumor size were analyzed using the Chi Square Test. Results: The Luminal A was the most common subtype of Indonesian BC (41.3%), followed by triple negative (25.5%), HER2 (19.4%) and luminal B (13.8%). Among the triple negative lesions, the basal-like subtype was more frequent than the non basal-like (58.8 % vs 41.2%). Luminal B accounted for the highest percentage of younger age cases (< 40 years old) while HER2+ was most common in older age (> 50 years old) patients. Triple negative/basal-like were commonly large in size. Age (p = 0.080) and tumor size (p = 0.462) were not significantly associated with molecular subtypes of BC. Conclusion: The most common molecular subtype of Indonesian BC is luminal A, followed by triple-negative, HER2+ and luminal B. The majority of triple-negative lesions are basal-like. There are no association between age and tumor size with molecular subtypes of Indonesian BCs. molecular subtypesbreast carcinomaagetumor size20180101161166http://journal.waocp.org/article_54073_6a84cbe6b2c8ea7692d40062c4ee6925.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Congenital Hypertrophy of Retinal Pigment Epithelium for Diagnosis of Familial Adenomatous Polyposis - the First FAP registry in IranSeyed KazemMirinezhadFaridehMousaviMasoodBagheriBitaSepehriAliGhavidelMortezaGhojazadehMohammad HossinSomi Objective: Familial adenomatous polyposis (FAP), an autosomal dominant inherited disorder is characterized by the presence of multiple adenomatous colorectal polyps, which can develop into cancer during early adulthood. Therefore, early diagnosis is essential. Most FAP patients have several extracolonic manifestations, including congenital hypertrophy of the retinal pigment epithelium (CHRPE). Whereas genetic markers may provide the main route to detection of ‘‘at risk’’ subjects , at present this approach is clearly limited and searches for a noninvasive phenotypic marker continue to be high priority.The aim of this study was to describe the pattern of distribution of CHRPE lesions and evaluate their diagnostic value in FAP patients and their family members in a local population. Methods: A total of 23 FAP patients and 26 relatives belonging to 12 families at high risk of developing FAP were subjected to colonoscopic and ophthalmological examination. Result: Retinal examinations demonstrated prevalences of CHRPE in FAP patents and their siblings of 78% and 38%, respectively. We were able to illustrate a significant correlation between FAP disease and the presence of retinal lesions. Sensitivity and specificity of CHRPE as a screening test to detect the presence of FAP are 78.3% and 61.5%, respectively, with a positive predictive value of 64.3% and a negative predictive value of 76.2 %. A "lesion form" significant difference was found between FAP and normal participants.Spearman nonparametric analysis revealed no correlation between age and number or size of lesions. Conclusion: Multiple CHRPE lesions are a diagnostic feature of FAP patients They are specific and sensitive clinical markers of this disease (specificity 60% and sensitivity 77%). Congenital Hypertrophy of the Retinal Pigment EpitheliumFamilial adenomatous polyposisColorectal CancerIran20180101167169http://journal.waocp.org/article_54076_3a4a1f951ee3adec2a9d2ff0a8c2c247.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Expression of VEGF and Cox-2 in Patients with Esophageal Squamous Cell CarcinomaCaio Cesar FlorianoLuzJulianaNogutiLeandroAraujoThiagoSimao GomesGianniMaraMarcelo De SouzaSilvaRicardoArtigiani Neto Esophageal cancer is a highly aggressive neoplasm. In Brazil, it is the sixth most frequent among men and fifteenth among women. The most common type is squamous cell carcinoma (SCC), responsible for 96% of cases. Twenty-eight specimens of Esophael squamous cell carcinoma (ESCC) were obtained by surgery procedures.The tissues were fixed in formalin and embedded in paraffin. In each case, all available hematoxylin and eosin stained sections were examined and a representative block was selected. The ages of these patients ranged from 40 to 93 years, with a mean age of 60 years. Results: The histological grade of tumors was 4 well-differentiated, 19 moderately differentiated and 5 poorly differentiated. Expression of Cox-2 and VEGF in ESCC was demonstrated in 23 (82,14%) and 13 (44,43%) cases, respectively. Adjacent normal mucosa was positive in 11 (39,29%) samples and 9 (32,15%) samples for Cox-2 and VEGF, respectively. No relationship between the expression of Cox-2 and VEGF with the clinicopathological parameters, including gender, age, surgical margin, lymph node status and tumor differentiation. The median follow-up period was 60 months. Survival analysis of patients with ESCC showed no relationship with the expression of Cox-2 and VEGF. Conclusion: VEGF and Cox-2 are expressed in ESCC. Cox-2, VEGF, play a significant role in the origin and development of ESCC and the inhibitors of these proteins could prove to be an important therapeutic tool in the control of this disease. Esophageal carcinomaCox-2VEGFprognosisimmunohistochemistry20180101171177http://journal.waocp.org/article_54009_897370e8fcc8d7e59b610b345017b6ac.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Can Histological Grade and Mitotic Index Replace Ki67 to Determine Luminal Breast Cancer Subtypes?DavidOddoDahianaPulgarNicoleElguetaFranciscoAcevedoDravnaRazmilicMaria ElenaNavarroMauricioCamusTomasMerinoIgnacioRetamalAlejandraPerez-SepulvedaAlejandraVillarroelHectorGalindoJoséPeñacesarsanchez Introduction: Breast cancer can be classified into subtypes based on immunohistochemical markers, with Ki67 expression levels being used to divide luminal BC tumors in luminal A and B subtypes; however, Ki67 is not routinely determined due to a lack of standardization. Objective: To evaluate histological grade and Eliminate: the mitotic index to determine if they can be used as an alternative method to Ki67 staining for luminal subtype definition. Methods: We evaluated estrogen receptor positive breast cancer tissue samples. Pathological analysis included determination of Ki67. A low level of Ki67 was defined as Breast neoplasmsKi67 antigenmitotic indexhistology20180101179183http://journal.waocp.org/article_55094_ada2e78b63233f99641fe5ef1d851eef.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Molecular Subtypes, Apoptosis and Proliferation Status in Indonesian Diffuse Large B-Cell Lymphoma CasesYosintaSnakIndrawatiFKartikaWidayatiNurArfianNungkiAnggorowati Objective: The diffuse large B-cell lymphoma (DLBCL) has two major molecular subtypes, germinal center B-cell-like (GCB) and non-GCB. These have differing behavior which affects overall patient survival. However, immunohistochemistry based molecular subtyping of Indonesian DLBCLs has been limited. This was the focus of the present study, with a focus of attention on the apoptotic index (AI) and the proliferation index (PI) of the two molecular subtypes. Materials and Methods: During the study period of 3.5 years, a total of 98 cases of DLBCL were identified. Molecular subtypes and PI were determined by immunohistochemistry and TUNEL method was used to determine the AI. Result: GCB accounted for 31 cases (31.6%) and non-GCB the remainder (68.4%). Gender showed a slight male predominance (54 cases, 55.1%), with a higher incidence in the extra-nodal region (57 cases, 58.2%). The AI and PI were significantly higher in GCB (pConclusion: The findings indicate that the non-GCB subtype is more common than GCB in Indonesian DLBCL. GCB features significantly higher PI and AI, which themselves appear linked. GCBnon-GCBproliferation indexapoptotic indexDLBCL20180101185191http://journal.waocp.org/article_55096_4cf4c75787d03179c2d6c9d6d51f3feb.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Evaluation of HPV DNA positivity in colorectal cancer patients in Kerman, Southeast IranRezaMalekpour AfsharZeinabDeldarHamid RezaMollaeiSeyed AlimohammadArabzadehMaryamIranpour Background: The HPV virus is known to be oncogenic and associations with many cancers has been proven. Although many studies have been conducted on the possible relationship with colorectal cancer (CRC), a definitive role of the virus has yet to be identified. Method: In this cross-sectional study, the frequency of HPV positivity in CRC samples in Kerman was assessed in 84 cases with a mean age of 47.7 ± 12.5 years over two years. Qualitative real time PCR was performed using general primers for the L1 region of HPV DNA. Results: Out of 84 CRC samples, 19 (22.6%), proved positive for HPV DNA. Genotyping of positive samples showed all of these to be of high risk HPV type. Prevalence of HPV infection appears to depend geographic region, life style, diet and other factors. Conclusion: In our location frequency of CRC is low, and this limited the sample size for evaluation of HPV DNA. The most prevalent types were HPV types 51 and 56. While HPV infection may play an important role in colorectal carcinogenesis, this needs to be assessed in future studies. Colorectal CancerHPVReal Time PCRHPV Genotype20180101193198http://journal.waocp.org/article_54329_c540a938552fdf4db10af4aba000ab92.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Genetic Polymorphism of Thiopurine S-methyltransferase in Children with Acute Lymphoblastic Leukemia in JordanMervatAlsousAl-MotassemYousefMariamAbdel JalilMohammedZawiahShorouqYacoubDeemaMomaniAliaGharabliSuhaOmarRawadRihani Background and Aims: It has been demonstrated that homozygote and heterozygote mutant allele carriers for thiopurine S-methyltransferase (TPMT) are at high risk of developing myelosuppression after receiving standard doses of 6-mercaptopurine (6-MP). The aim of this study was to determine the frequency of TPMT deficient alleles in children with acute lymphoblastic leukemia (ALL) in Jordan and to compare it with other ethnic groups. Methods: We included 52 ALL childhood cases from King Hussein Cancer Research Center in Jordan. Genotyping of the rs1800460, rs1800462, and rs1142345 SNPs was performed by polymerase chain reaction (PCR) followed by sequencing. Comparisons were made with historical data for controls and for both volunteers and cases from other middle-eastern countries. Results: Mutant TPMT alleles were present in 3.8% (2/52) of patients. Allelic frequencies were 1.0% for both TPMT*B and TPMT*C. None of the patients were heterozygous or homozygous for TPMT*3A or TPMT *2. We did not find statistically significant differences in the distribution of mutant alleles between Jordan and other middle-eastern countries for both healthy volunteers or ALL patients. Conclusions: The overall frequency of TPMT mutant alleles was low and did not exhibit differences compared to other middle-eastern countries, including Jordanian studies assessing TPMT mutant alleles in healthy volunteers. The current results question the value of TPMT genotyping in the Jordanian population. Keywords: thiopurine methytransferasechildhood acute lymphoblastic leukemiapharmacogenetics6-mercaptopurine20180101199205http://journal.waocp.org/article_54218_deededccaef853f94a45da05b7d2187f.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191RapidArc vs Conventional IMRT for Head and Neck Cancer Irradiation: Is Faster Necessary Better?KarimMashhourMahaKamaleldinWedadHashem Purpose: The aim of this study was to dosimetrically evaluate and compare double arc RapidArc (RA) with conventional IMRT (7 fields) plans for irradiation of locally advanced head and neck cancers (LAHNC), focusing on target coverage and doses received by organs at risk (OAR). Methods: Computed tomography scans of 20 patients with LAHNC were obtained. Contouring of the target volumes and OAR was done. Two plans were made for each patient, one using IMRT and the other double arc RA, and calculated doses to planning target volume (PTV) and OAR were compared. Monitor units for each technique were also calculated. Results: PTV coverage was similar with both techniques. The homogeneity index (HI) was higher for the IMRT plans with a value of 0.108 ± 0.021 compared to 0.0975 ± 0.017 for double arc RA plans (p-value of 0.540). The double arc RA plans achieved a better conformity with a CI95%= 1.01 ± 0.021 compared to 1.05 ± 0.057 achieved with the IMRT plans (p-value of 0.036). The average monitor units (MU) ±SD were 930.5 ± 142.42 for the IMRT plans as opposed to 484.25 ± 69.47 for the double arc RA plans (P-value of 0.002). Double arc plans provided better OAR sparing with a significant p-value of 0.002 and 0.004 for the right and left parotid glands, respectively. Conclusions: RA is a rapid and accurate technique that uses lower MUs than conventional IMRT. Double arc plans provide better dose conformity, OAR sparing and a more homogeneous target coverage compared to IMRT. dosimetricdouble arcIMRTMUs20180101207211http://journal.waocp.org/article_54331_80a4bd3e6edd91ca5bcfeef68b662de1.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Intra-Operative Frozen Sections for Ovarian Tumors – A Tertiary Center ExperienceNur ZaitiMd ArshadBeng KwangNgNoor AsmalizaMd PaimanZalehaAbdullah MahdyRushdanMohd Noor Background: Accuracy of diagnosis with intra-operative frozen sections is extremely important in the evaluation of ovarian tumors so that appropriate surgical procedures can be selected. Study design: All patients who with intra-operative frozen sections for ovarian masses in a tertiary center over nine years from June 2008 until April 2017 were reviewed. Frozen section diagnosis and final histopathological reports were compared. Main outcome measures: Sensitivity, specificity, positive and negative predictive values of intra-operative frozen section as compared to final histopathological results for ovarian tumors. Results: A total of 92 cases were recruited for final evaluation. The frozen section diagnoses were comparable with the final histopathological reports in 83.7% of cases. The sensitivity, specificity, positive predictive value and negative predictive value for benign and malignant ovarian tumors were 95.6%, 85.1%, 86.0% and 95.2% and 69.2%, 100%, 100% and 89.2% respectively. For borderline ovarian tumors, the sensitivity and specificity were 76.2% and 88.7%, respectively; the positive predictive value was 66.7% and the negative predictive value was 92.7%. Conclusion: The accuracy of intra-operative frozen section diagnoses for ovarian tumors is high and this approach remains a reliable option in assessing ovarian masses intra-operatively. Accuracyfrozen sectionovarian tumorsMalaysia20180101213218http://journal.waocp.org/article_54334_cb9b566823fb2b6b564df46d92a2c6f2.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular CarcinomasTarekAboushoushaSamahMamdouhHussamHamdyNohaHelalFatmaKhorshedGehanSafwatMohamedSeleem Objective: To investigate the expression of TTF-1, RAGE, GLUT1 and SOX2 in HCV-associated HCCs and in surrounding non-tumorous liver tissue. Material and Methods: Tissue material from partial hepatectomy cases for HCC along with corresponding serum samples and 30 control serum samples from healthy volunteers were studied. Biopsies were classified into: non-tumor hepatic tissue (36 sections); HCC (33 sections) and liver cell dysplasia (LCD) (15 sections). All cases were positive for HCV. Immunohistochemistry (IHC), gene extraction and quantitative real-time reverse-transcription assays (qRT-PCR) were applied. Results: By IHC, LCD and HCC showed significantly high percentages of positive cases with all markers. SOX2 showed significant increase with higher HCC grades, while RAGE demonstrated an inverse relation and GLUT-1 and TTF-1 lacked any correlation. In nontumorous-HCV tissue, we found significantly high TTF-1, low RAGE and negative SOX2 expression. RAGE, GLUT-1 and SOX2 show non-significant elevation positivity in high grade HCV compared to low grade lesions. TTF-1, RAGE and SOX2 exhibited low expression in cirrhosis compared to fibrosis. Biochemical studies on serum and tissue extracts revealed significant down-regulation of RAGE, GLUT-1 and SOX2 genes, as well as significant up-regulation of the TTF-1 gene in HCC cases compared to controls. All studied genes show significant correlation with HCC grade. In non-tumor tissue, only TTF-1 gene expression had a significant correlation with the fibrosis score. Conclusion: Higher expression of TTF-1, RAGE, GLUT-1 and SOX2 in HCC and dysplasia compared to non-tumor tissues indicates up-regulation of these markers as early events during the development of HCV-associated HCC. TTF1RAGEGLUT1SOX2HCC20180101219227http://journal.waocp.org/article_55065_fad95bde10c59df9f6bff1c00abb4638.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Minimum Residual Disease in Patients Post Radical Prostatectomy for Prostate Cancer: Theoretical Considerations, Clinical Implications and Treatment OutcomeNigel PMurraySocratesAedoCynthiaFuentealbaEduardoReyesAnibalSalazar Introduction: Minimal residual disease (MRD) remaining after curative therapy for prostate cancer has the potential for growth and can result in metastasis. Circulating prostate cells (CPCs) and bone marrow micro-metastasis (mM) could represent different types of MRD. We here determined; biochemical failure free survival rates; time to BF after 10 years follow-up; and the presence of CPCs and mM in patients treated with radical prostatectomy (RP) for prostate cancer. Methods and Patients: One month after RP, blood and bone marrow were sampled for assessment of CPCs and mM. Cases were classified as: group A, CPC negative and mM negative; group B, CPC negative and mM positive; Group C, CPC positive and mM negative; and Group D, CPC positive and mM positive. Subjects were followed with serial determination of PSA levels, recording the time at which BF occurred defined as a serum PSA >0.2ng/ml. After ten years of follow-up Kaplan-Meier survival curves were generated and the restricted mean survival time (RMST) for each group calculated. Results: A total of 321 men participated, 140 in group A with survival of 92.7% (86.3 to 96.2), 39 in group B with 55.8% (37.2 to 70.9); 54 in group C with 6.41% (1.19 to 18.21) and 88 in group D with 4.96%(1.64 to 11.13%. The RMST (in years) were: group A, 9.47 (9.24 to 9.69); group B, 9.23 (8.87 to 9.58); group C, 4.62 (4.46 to 4.77); and group D, 3.57 (3.52 and 3.63) (p-valueConclusions: CPC positive men have more aggressive disease, with increased early failure; men who are only positive for mM are at greater risk of late failure. These two forms of MRD represent different clinical entities with respect to biochemical failure and could be used to guide clinical treatment decisions. Prostate cancerminimal residual diseasebiochemical failurecirculating prostate cellsmicro-metastasis20180101229236http://journal.waocp.org/article_54337_39d88b1ec7ac97325ce53a0a421fe724.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Increased Risk of Penile Cancer among Men Working in AgriculturePorfírio Fernandes de MedeirosJuniorEugênio Henrique VilelaSilvaKevin LeiteMouraYasminde AquinoMathiasWeller Background: Previous Brazilian studies have indicated that working in agriculture could lead to an increased risk of penile cancer. The present descriptive study aimed at establishing a possible association between penile cancer and agricultural occupation. Materials and methods: Data on a total of 103 penile cancer patients were obtained from medical records of two reference centres for cancer treatment in the state of Paraíba, Northeast Brazil. Information about sexual behaviour was obtained in interviews for 48 cases. Results: Of 103 patients 38 and 52 were illiterate and had not completed graduation, respectively, and 60 earned less than twice the minimum wage. All together, 72 (70%) worked in agriculture and 39 confirmed involvement in application of agrochemicals. A history of phimosis was noted for 42 (69%) out of 61 patients and 40 (59%) out of 68 ever smoked. Pathological signs of HPV infection were detected in 45 (70%) out of 64 patients. Of the 48 interviewed patients, 27 (56%) confirmed sexual contact with prostitutes and eight (19%) out of 43 had sex with animals. Conclusions: Data confirmed the presence of several risk factors for penile cancer, like phimosis, smoking, HPV infection and promiscuous sexual behaviour. The high number of Brazilian agricultural workers with penile cancer was unexpected if compared with other professional groups. Future studies should focus on this group of men and elucidate possible reasons for their increased risk. Penile cancerEpidemiologyRisk factors20180101237241http://journal.waocp.org/article_53793_0a7be9ad859d677ed9e62bdcc775337d.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Cancer Diagnosis Epigenomics Scientific Workflow Scheduling in the Cloud Computing Environment Using an Improved PSO AlgorithmSadhasivamNBalamuruganRPandiM Objective: Epigenetic modifications involving DNA methylation and histone statud are responsible for the stable maintenance of cellular phenotypes. Abnormalities may be causally involved in cancer development and therefore could have diagnostic potential. The field of epigenomics refers to all epigenetic modifications implicated in control of gene expression, with a focus on better understanding of human biology in both normal and pathological states. Epigenomics scientific workflow is essentially a data processing pipeline to automate the execution of various genome sequencing operations or tasks. Cloud platform is a popular computing platform for deploying large scale epigenomics scientific workflow. Its dynamic environment provides various resources to scientific users on a pay-per-use billing model. Scheduling epigenomics scientific workflow tasks is a complicated problem in cloud platform. We here focused on application of an improved particle swam optimization (IPSO) algorithm for this purpose. Methods: The IPSO algorithm was applied to find suitable resources and allocate epigenomics tasks so that the total cost was minimized for detection of epigenetic abnormalities of potential application for cancer diagnosis. Result: The results showed that IPSO based task to resource mapping reduced total cost by 6.83 percent as compared to the traditional PSO algorithm. Conclusion: The results for various cancer diagnosis tasks showed that IPSO based task to resource mapping can achieve better costs when compared to PSO based mapping for epigenomics scientific application workflow. Cancer diagnosisGenomicsParticle Swam OptimizationScientific WorkflowScheduling20180101243246http://journal.waocp.org/article_53794_c50333ab45fc6483ce1ea6b5e1b47ceb.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Synthesis, Characterization, and Evaluation of Cancer Prevention Activity of Novel Modified Heterocyclic CompoundsVijayakumarKSountharrajanSSuganyaE Anticancer approaches may employ change of molecular structure to enhance preventive influence of chemical agents. The present examination concerned the potential anticancer impact of modified heterocyclic compounds. A strategy was developed to combine tetrazole moieties from different diamines with 2-aminobenzoic and substituted benzoyl chloride compounds with attention to synthesis, characterization and assessment of cancer preventive activity, applying IR, 1HNMR, 13CNMR and Mass spectra. CytotoxicityHeterocyclesMulticomponent reactions and sodiumazide20180101247252http://journal.waocp.org/article_55130_83f2b25fc3b671345123ccb856965152.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Dose Calculation Accuracy of AAA and AcurosXB Algorithms for Small Central and Interface Lung Lesions - Verification with Gafchromic Film DosimetryAnanda Giri BabuAlagarK.MGANESHKarunakaranKaviarasu Dose calculation for small field radiotherapy with heterogeneity often involves discrepancies, so that algorithms used by treatment planning systems (TPS) should be evaluated with reference to achieving optimal treatment results. Accuracy of two model based algorithms, AcurosXB (AcXB) and the analytical anisotropic algorithm (AAA) from Eclipse TPS, were here tested. Measurements are made using Gafchromic EBT3 films with indigenously generated lung phantoms irradiated with 6 MV photons. Lung phantoms contained two types of tumor plugs, one kept at an interface attached to the chest wall in right lung (RIT) and the other at the centre of the left lung (LCT). RIT and LCT were studied with two different tumor diameters, 1.5 cm and 2.5 cm. Scanned images were planned in TPS with 3D-CRT, IMRT and VMAT and individual plans for each tumor were irradiated keeping the Gafchromic film at the centre of the tumor to evaluate the dose distribution in the central plane. Both algorithms, irrespective of delivery techniques, showed more deviation with smaller than larger diameter tumors. Also, both demonstrated maximum deviation at the junction of tumor and lung in both RIT and LCT cases. However, the deviation observed was higher with AAA and a minimal acceptable deviation of within 4 % was achieved with AcurosXB. TPS AlgorithmsLung Heterogeneitydose plane20180101253259http://journal.waocp.org/article_55140_6a2aed9d63626407038d776452f4f2b2.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Fast Foods, Sweets and Beverage Consumption and Risk of Colorectal Cancer: A Case-Control Study in JordanReema FTayyemHiba ABawadiIhabShehadahKamal EBani-HaniHamedTakruriTareqAl-JaberiDennis DHeath Background: The effects of consuming fast foods, sweets and beverages on the development of colorectal cancer (CRC) are unclear. The aim of this case-control study was to assess possible associations between the consumption of different fast foods, sweets and beverages and CRC risk in a Jordanian population. Methods: Two hundred and twenty diagnosed CRC cases and 281 controls were enrolled. Diet history was obtained using a validated quantitative questionnaire. Results: Consumption of some types of fast food, and particularly falafel, was associated with an increased risk of developing CRC. Elevated risk was found for potato and corn chips with an AOR of 4.36 (95%CI: 1.24-15.28) for daily consumption and 3.33 (95%CI: 1.00-11.11) for ≥5 servings/week. Consuming 1-2 or >5 servings per week of fried potatoes or 2-3 servings per week of chicken in sandwiches also increased the risk while exposure to fresh tomato juice and hot pepper sauce on a monthly basis appeared to exert a protective effect. Conclusions: Consumption of fried fast food items was significantly linked with an increased risk of developing CRC in Jordan. Colorectal Cancerfast foodssweetsand beverages20180101261269http://journal.waocp.org/article_55145_0e0fd7d1707ca5502de0239f7d1c33fd.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Lack of Association between Red Meat Consumption and a Positive Fecal Immunochemical Colorectal Cancer Screening Test in Khon Kaen, Thailand: a Population- Based Randomized Controlled TrialPutthikraiPramualPongdechSarakarnSiripornKamsa-ardChananyaJirapornkulNaowaratManeeninPrasertThavondunstidPrachakJuntarachSupanneePromthet Background: There is convincing evidence from epidemiological studies that meat consumption increases colorectal cancer (CRC) risk. However, assessment of any association with a positive fecal immunochemical test (FIT) in CRC screening has been limited. If a link could be shown this might be helpful for establishing a risk group for colonoscopy. Objective: This study aimed to assess any association between meat consumption and other lifestyle factors and a positive FIT result in a Thai population. Methods: A cross-sectional analytical study was conducted with 1,167 participants in a population-based randomized controlled trial. CRC was screened from May 2016 - February 2017. Subjects aged 45-74 years who met the eligibility criteria were randomly allocated to the study arm. A positive FIT was determined with cut-off 100 ng/mL. Multiple logistic regression was used to analyze any relationship between lifestyle factors and a positive FIT. Result: The total number of subjects was 1,060 (90.8% return rate of FIT). With FIT100, FIT150, and FIT200, positive tests were found in 92 (8.68%), 74 (6.98%), and 60 (5.66%), respectively. No significant associations were noted with any of the variables, except for being aged 60-74 years (ORadj = 1.62, 95% CI: 1.03-2.54) Borderline significance was observed for high consumption of vegetables (ORadj = 0.62, 95% CI: 0.36-1.07) and being male (ORadj = 1.39, 95% CI: 0.87-2.22). Conclusion: Despite the evidence from the literature, no association was here found between a positive FIT result and meat consumption or other well-established lifestyle parameters. Being aged 60-74 years was a risk factor which should be taken into account in CRC screening strategy in countries like Thailand with limited access to endoscopy. Red meat consumptionfecal immunochemical testColorectal Cancer20180101271278http://journal.waocp.org/article_55305_78a21d876baae5954e8689c3c7079c07.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191Photodynamic Cancer Therapy Using Wavelet Based Monte Carlo Computation of Light Absorption with a Genetic AlgorithmMeenaakshi SundhariR P Objective: The method to treating cancer that combines light and light-sensitive drugs to selectively destroy tumour cells without harming healthy tissue is called photodynamic therapy (PDT). It requires accurate data for light dose distribution, generated with scalable algorithms. One of the benchmark approaches involves Monte Carlo (MC) simulations. This gives an accurate assessment of light dose distribution, but is very demanding in computation time, which prevents routine application for treatment planning. Methods: In order to resolve this problem, a design for MC simulation based on the gold standard software in biophotonics was implemented with a large modern wavelet based genetic algorithm search (WGAS). Result: The accuracy of the proposed method was compared to that with the standard optimization method using a realistic skin model. The maximum stop band attenuation of the designed LP, HP, BP and BS filters was assessed using the proposed WGAS algorithm as well as with other methods. Conclusion: In this paper, the proposed methodology employs intermediate wavelets which improve the diversification rate of the charged genetic algorithm search and that leads to significant improvement in design effort efficiency. Cancer therapySkin TreatmentsBiophotonicsWaveletTissue optical20180101279282http://journal.waocp.org/article_55138_bee6d55177e5b732cbf1bf4bc8224bd4.pdf2018-01-01Asian Pacific Journal of Cancer PreventionAsian Pac J Cancer Prev1513-73681513-73682018191A Modified Epirubicin and Oxaliplatin Plus Capecitabine (EOX) Regimen as a Second- Line Therapy in Patients with Advanced Gastric CancerYakupBozkayaNuriye YıldırımÖzdemirOzanYaziciNebi SerkanDemirciAlicanKurtipekGökmen UmutErdemYakupErgünNurullahZengin Objective: We aimed to evaluate the effectiveness of an mEOX (modified epirubicin, oxaliplatin plus capecitabine) regimen as second line therapy after failure of mDCF (modified docetaxel, cisplatin plus fluorouracil). Methods: Gastic cancer patients for whom first-line therapy was unsuccessful and who subsequently received mEOX (epirubicin 50 mg/ m2 on day 1, oxaliplatin 85 mg/m² day 1 and capecitabine twice-daily dose of 625 mg/ m2, p.o. for 2 weeks) every 3 weeks until disease progression or unacceptable toxicity, were retrospectively analyzed. Results: The study population comprised 129 cases with a median age of 55 years (range= 27-78), the majority being male (76 %). Most (75.2%) had ≥ 2 sites of metastasis. The median number of chemotherapy courses was five (range= 2–9). Forty-nine achieved a partial response and 33 showed stable disease, resulting in a ORR (overall response rate) of 38% and a DCR (disease control rate) of 63.6%. The most frequent features of grade 3-4 hematological and non-hematological toxicity were neutropenia (8.5%) and nausea/vomiting (5.4%). None of the patients suffered death due to toxicity. The median PFS was 4.7 months (95% CI, 4.1–5.3) and the OS was 7.4 months (95% CI, 6.3–8.5). On multivariate analysis, age ≥ 60 years and ECOG performance status (0-1) were independent prognostic factors affecting PFS and OS. Conslusions: In advanced gastric cancer patients, who progress after first line chemotherapy and have an ECOG performance status of 0-1, mEOX is a well tolerated triple regimen associated with a promising OS and PFS. Modified EOXGastric cancerModified DCFSecond-line Therapy20180101283290http://journal.waocp.org/article_54300_4ce583c6a496def77a6dbf3dfc706bca.pdf